LOCUS Exported 7034 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses N-terminus to aa270 of Cre recombinase fused to ABI CID. ACCESSION . VERSION . KEYWORDS pBW2661_pCAG-PV1-iCre-N270-L1-ABI-NLS-BGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7034) AUTHORS Weinberg BH, Cho JH, Agarwal Y, Pham NTH, Caraballo LD, Walkosz M, Ortega C, Trexler M, Tague N, Law B, Benman WKJ, Letendre J, Beal J , Wong WW TITLE High-performance chemical- and light-inducible recombinases in mammalian cells and mice JOURNAL Nature Communications (2019) volume 10, Article number: 4845 REFERENCE 2 (bases 1 to 7034) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7034 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..660 /label=chicken beta-actin promoter intron 661..1677 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1601..1623 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1685..1704 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 3555..3578 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 3579..3599 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 3609..3833 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4073..4092) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4138..4193 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4192..4211) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4554..4570) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4554..4570) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4567..4589) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4578..4594 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4602..4632) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4647..4668 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin 4753..4887 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4814..4833 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4907..5041 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4944..4963) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4998..5017 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5109..5126) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5280..5868) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5360..5379) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6039..6899) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6662..6681 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6900..7004) /gene="bla" /label=AmpR promoter ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct 421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga 481 tgggggcggg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg 541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt 601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg 661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg 721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct 781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa 841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc 901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc 961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg 1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg 1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac 1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt 1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg 1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc 1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg 1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc 1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag 1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg 1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg 1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc 1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt gattaattcg 1741 agcgaaggcg cgcccattac tcgcatccat tctcaggctg tctcgtctcg tctcgcacgc 1801 gtgccgccac catgtccaac ctgctgactg tgcaccaaaa cctgcctgcc ctccctgtgg 1861 atgccacctc tgatgaagtc aggaagaacc tgatggacat gttcagggac aggcaggcct 1921 tctctgaaca cacctggaag atgctcctgt ctgtgtgcag atcctgggct gcctggtgca 1981 agctgaacaa caggaaatgg ttccctgctg aacctgagga tgtgagggac tacctcctgt 2041 acctgcaagc cagaggcctg gctgtgaaga ccatccaaca gcacctgggc cagctcaaca 2101 tgctgcacag gagatctggc ctgcctcgcc cttctgactc caatgctgtg tccctggtga 2161 tgaggagaat cagaaaggag aatgtggatg ctggggagag agccaagcag gccctggcct 2221 ttgaacgcac tgactttgac caagtcagat ccctgatgga gaactctgac agatgccagg 2281 acatcaggaa cctggccttc ctgggcattg cctacaacac cctgctgcgc attgccgaaa 2341 ttgccagaat cagagtgaag gacatctccc gcaccgatgg tgggagaatg ctgatccaca 2401 ttggcaggac caagaccctg gtgtccacag ctggtgtgga gaaggccctg tccctggggg 2461 ttaccaagct ggtggagaga tggatctctg tgtctggtgt ggctgatgac cccaacaact 2521 acctgttctg ccgggtcaga aagaatggtg tggctgcccc ttctgccacc tcccaactgt 2581 ccacccgggc cctggaaggg atctttgagg ccacccaccg ctccggaggg tctggctccg 2641 gatcaagtgg tggcagcggt acccctttgt atggttttac ttcgatttgt ggaagaagac 2701 ctgagatgga agatgctgtt tcgactatac caagattcct tcaatcttcc tctggttcga 2761 tgttagatgg tcggtttgat cctcaatccg ccgctcattt cttcggtgtt tacgacggcc 2821 atggcggttc tcaggtagcg aactattgta gagagaggat gcatttggct ttggcggagg 2881 agatagctaa ggagaaaccg atgctctgcg atggtgatac gtggctggag aagtggaaga 2941 aagctctttt caactcgttc ctgagagttg actcggagat tgggtcagtt gcgccggaga 3001 cggttgggtc aacgtcggtg gttgccgttg ttttcccgtc tcacatcttc gtcgctaact 3061 gcggtgactc tagagccgtt ctttgccgcg gcaaaactgc acttccatta tccgttgacc 3121 ataaaccgga tagagaagat gaagctgcga ggattgaagc cgcaggaggg aaagtgattc 3181 agtggaatgg agctcgtgtt ttcggtgttc tcgccatgtc gagatccatt ggcgatagat 3241 acttgaaacc atccatcatt cctgatccgg aagtgacggc tgtgaagaga gtaaaagaag 3301 atgattgtct gattttggcg agtgacgggg tttgggatgt aatgacggat gaagaagcgt 3361 gtgagatggc aaggaagcgg attctcttgt ggcacaagaa aaacgcggtg gctggggatg 3421 catcgttgct cgcggatgag cggagaaagg aagggaaaga tcctgcggcg atgtccgcgg 3481 ctgagtattt gtcaaagctg gcgatacaga gaggaagcaa agacaacata agtgtggtgg 3541 tggttgattt gaaggattac aaggacgatg acgataagcc caagaaaaag cggaaggtgt 3601 gagaattcct gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 3661 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 3721 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 3781 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 3841 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcgtgtaccg 3901 ctgggagttc gtagacggaa acaaacgcag aatccaagcg cggccgccca ggatacatag 3961 attaccacaa ctccgagccc ttccaccgct agcgcactcc tcaggtgcag gctgcctatc 4021 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc 4081 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat 4141 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa 4201 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg 4261 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt 4321 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt 4381 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct 4441 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc 4501 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag 4561 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4621 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4681 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt 4741 cagcaaccat agtcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 4801 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 4861 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc 4921 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 4981 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat 5041 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 5101 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 5161 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 5221 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 5281 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 5341 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 5401 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 5461 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 5521 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 5581 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 5641 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 5701 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 5761 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 5821 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 5881 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 5941 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 6001 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 6061 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 6121 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 6181 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 6241 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 6301 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 6361 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 6421 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 6481 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 6541 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 6601 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 6661 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 6721 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 6781 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 6841 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 6901 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 6961 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 7021 gtgccacctg ggtc //